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目的 筛选与日本血吸虫尾蚴抗原有共同免疫原性的日本血吸虫成虫抗原分子 ,为血吸虫病诊断和疫苗研究提供新的候选分子。 方法 利用日本血吸虫尾蚴可溶性抗原免疫兔血清筛选日本血吸虫 (Schistosomajapon icum ,Sj)成虫cDNA文库 ,对阳性克隆的插入片段进行PCR扩增和序列分析。 结果 共筛选出 13个阳性克隆 ,PCR扩增出特异的插入片段。测序分析获 4个新基因SjCAI、SjCA、SjCAI 2和SjCAI 3 (登录号分别为AF495 883、AF5 15 83 4、AY1180 86和AY12 93 0 3 ) ,分别编码 3 5 3、161、13 7和 72个氨基酸的蛋白。SjCAI蛋白含 6个DNA结合锌指 ,与原肠胚锌指蛋白XLCGF48.2有一定同源性 ;SjCA蛋白、SjCAI 2蛋白和SjCAI 3蛋白含N 糖基化和磷酸化位点。 结论 筛选Sj成虫cDNA文库获得了新的基因序列。
Objective To screen adult schistosoma japonicum antigen molecules that share immunogenicity with Schistosoma japonicum cercariae antigen and provide new candidate molecules for the diagnosis and vaccine research of schistosomiasis. Methods Schistosoma japonicum soluble antigen was used to immunize rabbit serum to screen the adult Schistosoma japonicum (Sj) cDNA library. The inserted fragment of positive clone was amplified by PCR and sequenced. Results A total of 13 positive clones were screened and specific insertions were amplified by PCR. Four new genes, SjCAI, SjCA, SjCAI 2 and SjCAI 3 (accession numbers AF495 883, AF5 15 83 4, AY1180 86 and AY12 93 0 3, respectively) were obtained by sequencing analysis and were coded as 3 5 3,161,13 7 and 72 respectively Amino acid protein. SjCAI protein contains 6 DNA-binding zinc fingers and has some homology with gastrula zinc finger protein XLCGF48.2; SjCA protein, SjCAI 2 protein and SjCAI 3 protein contain N-glycosylation and phosphorylation sites. Conclusion A new gene sequence was obtained by screening Sj adult cDNA library.